Radioactive measurement of 2′,3′-cyclic nucleotide 3′-phosphodiesterase activity in the central and peripheral nervous system and in extraneural tissue
| Autor: | G.M. McKhann, T. J. Sprinkle, M. E. Zaruba |
|---|---|
| Rok vydání: | 1978 |
| Předmět: |
Biophysics
Extraneural Biochemistry 2' 3'-Cyclic-nucleotide 3'-phosphodiesterase Hydrolysis Column chromatography Isomerism Cyclic AMP medicine Animals Molecular Biology Chromatography Phosphoric Diester Hydrolases Chemistry Brain Substrate (chemistry) Cell Biology Sciatic Nerve Adenosine Rats medicine.anatomical_structure Isotope Labeling Peripheral nervous system Alkaline phosphatase 2' 3'-Cyclic-Nucleotide Phosphodiesterases medicine.drug |
| Zdroj: | Analytical Biochemistry. 88:449-456 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/0003-2697(78)90443-8 |
| Popis: | This paper describes a new, sensitive, and reproducible method for the determination of 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (EC 3.1.4.37) activity in both central and peripheral nervous system tissue, as well as in extraneural tissue. Radioactive [8- 3 H]adenosine 2′,3′-cyclic monophosphate was used as the substrate. The [8- 3 H]2′-AMP product formed was isolated by thin-layer chromatography, or, alternatively, the reaction was coupled with an excess of Escherichia coli alkaline phosphatase, and the [8- 3 H]adenosine formed was isolated by column chromatography on AG 1-X2 resin. The values obtained by the two methods were compared with those obtained using a spectrophotometric method. Hydrolysis rates of 0.50 nmol/min could be reproducibly measured in 18-day fetal rat spinal cord. |
| Databáze: | OpenAIRE |
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